您好，欢迎访问上海市农业科学院 机构知识库！
上海市农业科学院机构知识库

全部

  • 全部
  • 标题
  • 学者
  • 机构
  • 关键词

Validation of a cotton-specific gene, Sad1, used as an endogenous reference gene in qualitative and real-time quantitative PCR detection of transgenic cottons

文献类型： 外文期刊

作者： Yang, LT 1 ; Chen, JX 2 ; Huang, C 2 ; Liu, YH 3 ; Jia, SR; Pan, LW; Zhang, DB;

作者机构： 1.Shanghai Acad Agr Sci, Agrobiotech Res Ctr, Key Lab Agr Genet & Breeding, Shanghai 201106, Peoples R China

2.Shanghai Acad Agr Sci, Agrobiotech Res Ctr, Key Lab Agr Genet & Breeding, Shanghai 201106, Peoples R China; Nanjing Univ, Dept Biol Sci & Technol, Nanjing 210093, Peoples R China; Shanghai Jiao Tong Univ, Sch Life Sci & Biotechnol, Shanghai 200240, Peoples R China; Chinese Acad Agr Sci, Biotechnol Res Inst, Beijing 100081, Peoples R China; Shanghai Entry Exit Inspect & Quarantine Bur, Shanghai 200070, Peoples R China

3.Shanghai Acad Agr Sci, Agrobiotech Res Ctr, Key Lab Agr Genet & Breeding

关键词： Gossypium hirsutum;stearoyl-acyl carrier protein desaturase gene;endogenous reference gene;genetically modified organism;conventional and real-time PCR

期刊名称：PLANT CELL REPORTS （ 影响因子：4.57； 五年影响因子：4.463 ）

ISSN： 0721-7714

年卷期： 2005 年 24 卷 4 期

页码：

收录情况： SCI

摘要： Genetically modified (GM) cotton lines have been approved for commercialization and widely cultivated in many countries, especially in China. As a step towards the development of reliable qualitative and quantitative PCR methods for detecting GM cottons, we report here the validation of the cotton (Gossypium hirsutum) endogenous reference control gene, Sad1, using conventional and real-time (RT)-PCR methods. Both methods were tested on 15 different G. hirsutum cultivars, and identical amplicons were obtained with all of them. No amplicons were observed when DNA samples from three species of genus Gossypium, Arabidopsis thaliana, maize, and soybean and others were used as amplified templates, demonstrating that these two systems are specific for the identification and quantification of G. hirsutum. The results of Southern blot analysis also showed that the Sad1 gene was two copies in these 15 different G. hirsutum cultivars. Furthermore, one multiplex RT-quantitative PCR employing this gene as an endogenous reference gene was designed to quantify the Cry1A(c) gene modified from Bacillus thuringiensis (Bt) in the insect-resistant cottons, such as Mon531 and GK19. The quantification detection limit of the Cry1A(c) and Sad1 genes was as low as 10 pg of genomic DNA. These results indicat that the Sad1 gene can be used as an endogenous reference gene for both qualitative and quantitative PCR detection of GM cottons.

  • 相关文献

[1]Screening and construct-specific detection methods of transgenic Huafan No 1 tomato by conventional and real-time PCR. Yang, LT,Shen, HF,Pan, AH,Chen, JX,Huang, C,Zhang, DB. 2005

[2]Validation of a rice specific gene, sucrose phosphate synthase, used as the endogenous reference gene for qualitative and real-time quantitative PCR detection of transgenes. Ding, JY,Jia, JW,Yang, LT,Wen, HB,Zhang, CM,Liu, WX,Zhang, DB. 2004

[3]Identification and Quantification of Genetically Modified Moonshade Carnation Lines Using Conventional and TaqMan Real-Time Polymerase Chain Reaction Methods. Li, Peng,Jia, Junwei,Bai, Lan,Pan, Aihu,Tang, Xueming,Li, Peng,Jia, Junwei,Bai, Lan,Pan, Aihu,Tang, Xueming. 2013

作者其他论文 更多>>
置顶
购物车
反馈

© 京ICP备09089781号-29 主办单位：上海市农业科学院

学术资源： 中国农业科学院 农业专业知识服务系统 农科联盟资源共建共享平台 中国农业科技文献与信息服务平台 中国农业期刊集成服务平台